PLGA-loaded puerarin attenuates arsenic-induced liver injury through modulation of ferroptosis, oxidative stress, inflammation, and apoptotic signaling.

BACKGROUND: Arsenic (AR) is an environmental and mining occupational toxicant with reported hepatotoxic effects. Ferroptosis has recently emerged as a critical mechanism involved in arsenic-induced hepatotoxicity. This study investigated the hepatoprotective effects of puerarin (PU) and its PLGA-loaded nanoparticle formulation (PU-PLGA) against arsenic-induced liver injury in rats, with particular emphasis on ferroptosis pathways modulation. METHODS: Sixty adult male Sprague-Dawley rats were randomly assigned into six groups: control, PU, PU-PLGA, arsenic (AR), AR + PU, and AR + PU-PLGA. Subacute hepatotoxicity was induced by oral administration of arsenic (10 mg/kg) for 14 days. Oxidative stress, ferroptosis-related biomarkers, inflammatory mediators, apoptotic markers, fibrotic indicators, DNA damage, histopathological alterations, and ultrastructural changes were evaluated. RESULTS: Arsenic exposure induced marked hepatic injury characterized by Fe²⁺ accumulation, increased lipid peroxidation markers (MDA and 4-HNE), upregulation of ACSL4, and suppression of GPX4 and SLC7A11, indicating enhanced ferroptotic activity. These alterations were accompanied by oxidative stress, inflammatory activation (NF-κB, TNF-α, IL-6, IL-1β, and COX-2), apoptotic signaling (Bax and caspase-3), DNA damage (8-OHdG and DNA fragmentation), and fibrotic responses (TGF-β and collagen I). Treatment with PU and PU-PLGA significantly attenuated arsenic-induced hepatic injury by reducing iron accumulation and lipid peroxidation, restoring GPX4 and SLC7A11 expression, and suppressing ACSL4 upregulation. Notably, PU-PLGA demonstrated greater efficacy than free PU in modulating ferroptosis-related biomarkers, restoring antioxidant and survival signaling (Nrf2, p-AKT, and Bcl-2), and improving hepatic histopathological and ultrastructural integrity. CONCLUSION: P PU-PLGA nanoparticles exerted potent hepatoprotective effects against arsenic-induced liver injury, associated with modulation of ferroptosis-related pathways, oxidative stress, inflammatory responses, apoptosis-associated signaling, and profibrotic alterations. These findings highlight PU-PLGA as a promising nanotherapeutic strategy for arsenic-associated hepatotoxicity, although further mechanistic and translational validation is warranted.